内毒素刺激Kupffer细胞对肝星状细胞前胶原基因表达的影响  

Effect of Endotoxin-stimulated Kupffer Cells on Procollagen Gene Expression in Hepatic Stellate Cells

作　　者：贾晋斌 吴晗 杨东仁[2] 陆伦根[3] 韩德五[4] 

机构地区：[1]康哲医药研究(深圳)有限公司,518057 [2]美国加州大学戴维斯分校分子生物学系 [3]上海交通大学附属第一人民医院消化科 [4]山西医科大学肝病研究所

出　　处：《胃肠病学》2011年第8期459-463,共5页Chinese Journal of Gastroenterology

摘　　要：背景:肠源性内毒素血症对肝纤维化具有启动作用,肝星状细胞(HSC)活化是肝纤维化发生、发展的中心环节。目的:探讨内毒素刺激Kupffer细胞对HSC前胶原基因表达的影响及其可能机制。方法:改良链酶蛋白酶、胶原酶原位灌流和密度梯度离心分离大鼠肝脏Kupffer细胞和HSC。以RNA斑点杂交检测脂多糖(LPS)、Kupffer细胞、LPS与Kupffer细胞共培养和肿瘤坏死因子-α(TNF-α)对HSC前胶原mRNA表达的影响,放射免疫法检测LPS对Kupffer细胞和HSC产生TNF-α的影响,RNA印迹法检测LPS和TNF-α对Kupffer细胞和HSC转化生长因子-β(TGF-β)mRNA表达的影响结果:经低浓度(0.5、1、5μg/ml)LPS处理的Kupffer细胞培养上清可增加HSCⅠ、Ⅲ、Ⅳ型前胶原mRNA表达,经高浓度(10、15、20μg/ml)LPS处理的Kupffer细胞培养上清、单独LPS、未经处理的Kupffer细胞培养上清、单独TN-α均无此作用。Kupffer细胞培养上清中的TNF-α含量随LPS浓度梯度的增加而递增,HSC培养上清中的TNF-α含量则无明显变化。TNF-α不能增加HSC TGF-βmRNA表达,但能增加Kupffer细胞TGF-βmRNA表达;经LPS或TNF-α处理的Kupffer细胞培养上清能增加HSC TGF-βmRNA表达。结论:低浓度内毒素能上调HSC前胶原基因表达,该作用有赖于内毒素激活Kupffer细胞所产生的活性介质的参与。Background：Intestinal endotoxemia can initiate liver fibrosis,and activation of hepatic stellate cells（HSC） is the central event in liver fibrosis.Aims：To investigate the effect of endotoxin-stimulated Kupffer cells（KC） on procollagen gene expression in HSC and its possible mechanism.Methods：KC and HSC were isolated from liver of rat by modified in situ perfusion with pronase and collagenase and density gradient centrifugation.RNA dot blot hybridization was used to detect the effect of lipopolysaccharide（LPS）,unstimulated KC-conditioned medium（KCCM）,LPS-activated KCCM and tumor necrosis factor-α（TNF-α） on procollagen mRNA expression in HSC;radioimmunoassay was used to detect the effect of LPS on TNF-αproduction in KC and HSC;and Northern blot was used to detect the effect of LPS and TNF-αon transforming growth factor-β（TGF-β） mRNA expression in KC and HSC.Results：KCCM treated with low-dose LPS（0.5, 1,5μg/ml） could enhance typeⅠ,Ⅲ,andⅣprocollagen mRNA expressions in HSC,while KCCM treated with high-dose LPS（10,15,20μg/ml）,LPS alone,unstimulated KCCM and TNF-αalone had no such effect.With the increase of LPS concentration,TNF-αwas parallelly increased in KCCM but no significant change was seen in HSC conditioned medium.TNF-αcould enhance TGF-βmRNA expression in KC but not in HSC;KCCM treated with either LPS or TNF-αcould enhance TGF-βmRNA expression in HSC.Conclusions：Low-dose endotoxin can enhance the expression of procollagen gene in HSC,which may depend on the involvement of active media produced by endotoxin-stimulated KC.

关 键 词：内毒素类 肿瘤坏死因子α 转化生长因子β 枯否细胞 肝星状细胞 前胶原 

分 类 号：R575.2[医药卫生—消化系统]